Bartonella Alsatica Research Articles

Prevalence and genetic diversity of rodent-associated Bartonella in Hulunbuir border regions, China

Bartonella spp. are globally distributed gram-negative facultative intracellular bacteria that infect a wide range of hosts. Rodents are natural reservoirs of many Bartonella species, some of which are also pathogenic to humans. The rapid development of transportation and tourism has highlighted the risk of Bartonella transmission to humans. Thus, it is essential to maintain surveillance of Bartonella spp. infections in rodents. In China, Bartonella spp. infections have been monitored in various areas; however, these have not included the Hulunbuir border regions. In the present study, we monitored the prevalence and genetics of rodent-associated Bartonella spp. in the Hulunbuir border regions. Eleven rodent species were captured at five ports. Eight species were confirmed as Bartonella-positive using quantitative PCR assay, with an overall positivity rate of 20.05 %. Lasiopodomys brandtii was the predominant rodent species captured for Bartonella detection. Sequencing and phylogenetic analysis (using the maximum likelihood method) revealed the presence of three Bartonella species in these rodents, including two pathogenic to humans, namely, Bartonella alsatica and Bartonella grahamii. B. grahamii was the predominant Bartonella species identified in the rodents. Taken together, these results highlight the prevalence and genetic diversity of Bartonella spp. in rodents in the Hulunbuir border regions, indicating the need for risk assessment of human spillover.

Bartonella species bacteremia in association with adult psychosis.

The potential role of pathogens, particularly vector-transmitted infectious agents, as a cause of psychosis has not been intensively investigated. We have reported a potential link between Bartonella spp. bacteremia and neuropsychiatric symptoms, including pediatric acute onset neuropsychiatric syndrome and schizophrenia. The purpose of this study was to further assess whether Bartonella spp. exposure or infection are associated with psychosis. In a blinded manner, we assessed the presence of anti-Bartonella antibodies by indirect immunofluorescence assays (IFA), and infection by amplification of bacterial DNA from blood by quantitative polymerase chain reaction (qPCR), digital PCR (dPCR), and droplet digital PCR (ddPCR) in 116 participants. Participants were categorized into one of five groups: 1) controls unaffected by psychosis (n = 29); 2) prodromal participants (n = 16); 3) children or adolescents with psychosis (n = 7); 4) adults with psychosis (n = 44); and 5) relatives of a participant with psychosis (n = 20). There was no significant difference in Bartonella spp. IFA seroreactivity between adults with psychosis and adult controls unaffected by psychosis. There was a higher proportion of adults with psychosis who had Bartonella spp. DNA in the bloodstream (43.2%) compared to adult controls unaffected by psychosis (14.3%, p = 0.021). The Bartonella species was determined for 18 of the 31 bacteremic participants, including infection or co-infection with Bartonella henselae (11/18), Bartonella vinsonii subsp. berkhoffii (6/18), Bartonella quintana (2/18), Bartonella alsatica (1/18), and Bartonella rochalimae (1/18). In conjunction with other recent research, the results of this study provide justification for a large national or international multi-center study to determine if Bartonella spp. bacteremia is more prevalent in adults with psychosis compared to adults unaffected by psychosis. Expanding the investigation to include a range of vector-borne and other microbial infections with potential CNS effects would enhance knowledge on the relationship between psychosis and infection.

Bartonella alsatica in Wild and Domestic Rabbits (Oryctolagus cuniculus) in The Netherlands

Members of the genus Bartonella are Gram-negative facultative intracellular bacteria that are transmitted by arthropod vectors. Bartonella alsatica was detected in the spleens and livers of 7 out of 56 wild rabbits (Oryctolagus cuniculus) and in the liver of 1 out of 87 domestic rabbits in the Netherlands. The molecular evidence of B. alsatica infection in wild as well as domestic rabbits indicates the possibility of exposure to humans when these come in close contact with rabbits and possibly their fleas with subsequent risk of Bartonella infection and disease.

Complete Genome Sequence of Bartonella alsatica Strain IBS 382 (CIP 105477).

Bartonella alsatica causes bacteremia in rabbits and, rarely, human infections. Here, we announce the complete and closed genome of B. alsatica IBS 382 (CIP 105477), generated by long-read Pacific Biosciences single-molecule real-time (SMRT) sequencing. The availability of this genome sequence allows future work on understanding the zoonotic potential of this pathogen.

Longitudinal Study of Bacterial Infectious Agents in a Community of Small Mammals in New Mexico.

Background and Objectives: Vector-borne bacterial diseases represent a substantial public health burden and rodents have been recognized as important reservoir hosts for many zoonotic pathogens. This study investigates bacterial pathogens in a small mammal community of the southwestern United States of America. Methods: A total of 473 samples from 13 wild rodent and 1 lagomorph species were tested for pathogens of public health significance: Bartonella, Brucella, Yersinia, Borrelia, Rickettsia spp., and Anaplasma phagocytophilum. Results: Three animals were positive for Yersinia pestis, and one Sylvilagus audubonii had a novel Borrelia sp. of the relapsing fever group. No Brucella, Rickettsia, or A. phagocytophilum infections were detected. Bartonella prevalence ranged between 0% and 87.5% by animal species, with 74.3% in the predominant Neotoma micropus and 78% in the second most abundant N. albigula. The mean duration of Bartonella bacteremia in mark-recaptured N. micropus and N. albigula was 4.4 months, ranging from <1 to 18 months, and differed among Bartonella genogroups. Phylogenetic analysis of the Bartonella citrate synthase gene (gltA) revealed 9 genogroups and 13 subgroups. Seven genogroups clustered with known or previously reported Bartonella species and strains while two were distant enough to represent new Bartonella species. We report, for the first time, the detection of Bartonella alsatica in North America in Sylvilagus audubonii and expand the known host range of Bartonella washoensis to include Otospermophilus variegatus. Interpretation and Conclusion: This work broadens our knowledge of the hosts and geographic range of bacterial pathogens that could guide future surveillance efforts and improves our understanding of the dynamics of Bartonella infection in wild small mammals.

Pathogens in fleas collected from cats and dogs: distribution and prevalence in the UK

BackgroundFleas (Siphonaptera) are the most clinically important ectoparasites of dogs and cats worldwide. Rising levels of pet ownership, climate change and globalisation are increasing the importance of a detailed understanding of the endemicity and prevalence of flea-borne pathogens. This requires continued surveillance to detect change. This study reports a large-scale survey of pathogens in fleas collected from client-owned cats and dogs in the UK.MethodsRecruited veterinary practices were asked to follow a standardised flea inspection protocol on a randomised selection of cats and dogs brought into the practice in April and June 2018. A total of 326 practices participated and 812 cats and 662 dogs were examined. Fleas were collected, identified to species and pooled flea samples from each host were analysed for the presence of pathogens using PCR and sequence analysis.ResultsOverall, 28.1% of cats and 14.4% of dogs were flea infested. More than 90% of the fleas on both cats and dogs were cat fleas, Ctenocephalides felis felis. Fleas of the same species from each infested host were pooled. DNA was amplified from 470 of the pooled flea samples using conventional PCR, 66 of which (14% ± 95% CI 3.14%) were positive for at least one pathogen. Fifty-three (11.3% ± 95% CI 2.85%) of the pooled flea DNA samples were positive for Bartonella spp., 35 were from cats and 4 from dogs, the remainder had no host record. Seventeen of the Bartonella spp. samples were found to be Bartonella henselae, 27 were Bartonella clarridgeiae (of two different strains), 4 samples were Bartonella alsatica and one was Bartonella grahamii; 4 samples could not be identified. Fourteen (3% ± 95% CI 1.53%) of the flea DNA samples were found to be positive for Dipylidium caninum, 10 of the D. caninum-infected samples were collected from cats and one from a dog, the other 3 positive flea samples had no host species record. Only 3 flea samples were positive for Mycoplasma haemofelis or Mycoplasma haemocanis; 2 were collected from cats and one had no host species record. Three fleas were positive for both D. caninum and Bartonella spp. One flea was positive for both Bartonella spp. and M. haemofelis or M. haemocanis.ConclusionsThis study highlights the need for ongoing flea control, particularly given the relatively high prevalence of Bartonella spp., which is of concern for both animal welfare and human health. The study demonstrates the ongoing need to educate pet owners about the effects of both flea infestation and also the pathogen risks these fleas present.

Genetic Diversity of Bartonella spp. in Wild Mammals and Ectoparasites in Brazilian Pantanal.

The present work aimed to investigate the genetic diversity of Bartonella in mammals and ectoparasites in Pantanal wetland, Brazil. For this purpose, 31 Nasua nasua, 78 Cerdocyon thous, 7 Leopardus pardalis, 110 wild rodents, 30 marsupials, and 42 dogs were sampled. DNA samples were submitted to a quantitative real-time PCR assay (qPCR). Positive samples in qPCR were submitted to conventional PCR assays targeting other five protein-coding genes. Thirty-five wild rodents and three Polygenis (P.) bohlsi bohlsi flea pools showed positive results in qPCR for Bartonella spp. Thirty-seven out of 38 positive samples in qPCR were also positive in cPCR assays based on ftsZ gene, nine in nuoG-cPCR, and six in gltA-cPCR. Concatenated phylogenetic analyses showed that two main genotypes circulate in rodents and ectoparasites in the studied region. While one of them was closely related to Bartonella spp. previously detected in Cricetidae rodents from North America and Brazil, the other one was related to Bartonella alsatica, Bartonella pachyuromydis, Bartonella birtlesii, Bartonella acomydis, Bartonella silvatica, and Bartonella callosciuri. These results showed that at least two Bartonella genotypes circulate among wild rodents. Additionally, the present study suggests that Polygenis (P.) bohlsi bohlsi fleas could act as possible Bartonella vectors among rodents in Pantanal wetland, Brazil.

Detection of Bartonella alsatica in European wild rabbit and their fleas (Spilopsyllus cuniculi and Xenopsylla cunicularis) in Spain.

BackgroundBartonella alsatica has been formerly isolated from the blood of wild European rabbit (Oryctolagus cuniculus) and identified as causative agent of human endocarditis and lymphadenitis. Fleas are known biological vectors for Bartonella sp. This report details the specific detection of B. alsatica in three flea species commonly associated with the European wild rabbit in Southern Iberian Peninsula (Odontopsyllus quirosi, Spylopsyllus cuniculi and Xenopsylla cunicularis).MethodsIn the present study we have tested the presence of Bartonella alsatica in 26 European wild rabbit specimens and the fleas that they carrying at the moment of capture. Together to rabbits, captured from different localities of Andalusia (Jaen, Granada and Cordoba provinces), we evaluated three of fleas species that parasitize it usually using molecular techniques [PCR amplification and sequencing of intergenic transcribed spacer (ITS) 16S-23S rRNA].ResultsOver a sample of 26 wild rabbits carrying fleas, positive PCR amplicons for B. alsatica were obtained from 10 rabbits. All positive flea pools for B. alsatica were collected from positive rabbits [33.33% (8/24 pools) of S. cuniculi, 33.33% (5/15 pools) of X. cunicularis and 0% (0/7 pools) of O. quirosi]. In three positive rabbits, a pool of S. cuniculi and two pools of X. cunicularis respectively were negative. After sequencing, only B. alsatica (Genbank accession AF312506) was found in the rabbits sampled as well as in S. cuniculi and X. cunicularis within the respective fleas.ConclusionsThis research confirms the implication of two pulicidae flea species, S. cuniculi and X. cunicularis in the maintenance of infection by B. alsatica in wild rabbit populations throughout the year. The zoonotic character of this bartonellosis emphasizes the need to alert public health authorities and the veterinary community for the risk of infection.

Molecular Detection ofBartonella alsaticain Rabbit Fleas, France

To the Editor: Bartonella alsatica was first isolated from the blood of wild rabbits from the Alsace region in France (1). This bacterium is now considered an emerging infectious disease zoonotic agent in persons in close contact with rabbits; at least 2 human cases of endocarditis and 1 human case of lymphadenitis have been reported (2–4). In this study, we report the molecular detection of B. alsatica in fleas (Spilopsyllus cuniculi) collected from rabbits in southern France. During January and February 2008, a total of 60 fleas were collected from wild rabbits (Oryctolagus cuniculus) from 3 regions in southern France: Canohes (42°38′N, 2°51′E), Pollestres (42°38′N, 2°52′E), and Toreilles (42°45′N, 2°58′E). The fleas were collected and identified phenotypically, kept in ethanol, and sent to Unite de Recherche sur les Maladies Infectieuses et Tropicales Emergentes in September 2009. DNA from these fleas, as well as negative controls from uninfected lice maintained as colonies in our laboratory, were extracted by using a QIAmp Tissue Kit (QIAGEN, Hilden, Germany), as described (5). Identification of flea species at the molecular level was achieved by PCR amplification and sequencing of partial siphonapteran 18S rDNA gene (1.95 kbp) as described (5). Sequences were assembled in Sequencher 4.2 (GeneCodes Corporation, Ann Arbor, MI, USA). DNA was used as templates in a real-time quantitative PCR specific for a portion of the Bartonella genus 16S–23S intergenic spacer (ITS) performed in a Smart cycler instrument (Cepheid, Sunnyvale, CA, USA), as described (2). Positive samples at the genus level were confirmed by PCR amplification and sequencing of the Bartonella ITS region, as described (2). Finally, B. alsatica amplification and specific identification was confirmed by using 2 new specific PCRs with primers and TaqMan probes (Applied Biosystems, Courtaboeuf, France) specific for a portion of the heat shock protein 60 (hsp60) and the DNA gyrase subunit B (gyrB) genes of B. alsatica (Table). Specificity of these 2 PCRs was verified in silico (computer simulation) and by using a panel of 14 Bartonella species available in our laboratory (data not shown). Table Oligonucleotide primers and TaqMan* fluorescent probe sequences of hsp60 and gyrB genes used for reverse transcription PCRs of Bartonella alsatica† All fleas were morphologically identified as S. cuniculi by using current taxonomic criteria (6). Moreover, the 18S rRNA gene amplified and sequenced as described (6) from fleas gave a sequence with 100% similarity with the sequence of S. cuniculi fleas deposited in GenBank (accession no. {type:entrez-nucleotide,attrs:{text:EU336097,term_id:169248332,term_text:EU336097}}EU336097). B. alsatica was detected by ITS reverse transcription–PCR in 8 (13.3%) of 60 fleas: 6 from Toreilles (17.6%, 6/34) region, 2 from Canohes (10.5%; 2/19), and none from Pollestres (0/7). Sequences obtained after PCR amplification and sequencing of partial ITS showed 96.6% identity with B. alsatica (GenBank accession no. {type:entrez-nucleotide,attrs:{text:HM060955,term_id:296881361,term_text:HM060955}}HM060955). Using our 2 new PCRs specific for partial hsp60 and gyrB genes from B. alsatica, we identified all Bartonella spp.–positive fleas, which had cycle threshold values ranging from 12.15 to <32.35 and 13.21 to <36.99 for hsp60 and gyrB genes, respectively. We report the specific detection of B. alsatica in S. cuniculi rabbit fleas from southern France using 4 different PCRs and sequencing, including 2 new reverse transcription PCRs described in this study. There is 1 report of molecular detection of B. alsatica from S. cuniculi fleas from a European wildcat (Felis silvestris silvestris) in Andalusia, Spain (7). Although S. cuniculi fleas are rare on cats, this study demonstrates that cats in contact with rabbits may be infected by these fleas and consequently become a potential source for B. alsatica transmission to humans. Marquez has also recently reported the molecular detection of B. alsatica in blood from 48/279 (17.2%) of wild rabbits (O. cuniculus) in Andalusia, Spain (8). In conclusion, further research is needed to better understand the mode of transmission of B. alsatica in humans and mammals and the role of rabbit fleas for potential transmission for these bacteria. The recent description of B. alsatica as a human pathogen and the discovery of rabbit fleas as a potential vector reemphasize the emergence potential of this bacterium in humans who have close contact with rabbits.

Molecular Detection of Bartonella alsatica in European Wild Rabbits (Oryctolagus cuniculus) in Andalusia (Spain)

A sample of 279 European wild rabbits, Oryctolagus cuniculus (141 males, 138 females), captured alive in Andalusia (Spain) and belonging to the two haplotype classes previously described for this species (230 and 49 corresponding with haplotypes A and B, respectively), were tested for the presence of Bartonella alsatica DNA. Two species-specific nested polymerase chain reaction assays targeting for 16S-23S rRNA intergenic spacer region and RNA polymerase β subunit genes have been developed. Forty-eight (17.20%) rabbits were infected with B. alsatica. Two-way contingency table analyses and the calculation of Cramer's V statistic showed no differences in infection rate, considering haplotype lineage or sex. The risk of infection of human population, especially for hunters in close contact with this demonstrated human pathogen, should be considered.

Bartonella alsatica endocarditis in a French patient in close contact with rabbits

Bartonella species are Gram-negative bacilli that belong to the alpha 2 subgroup of Proteobacteria and are agents of blood culture-negative endocarditis (BCNE) [1]. Among the Bartonella species involved in BCNE, B. henselae and B. quintana are the main causative agents [1]. However, endocarditis due to other rare Bartonella spp. have been reported, including B. elizabethae, B. vinsonii subsp. berkhoffii, B. vinsonii subsp. arupensis, B. koehlerae and recently B. alsatica [2,3]. B. alsatica is a recently identified agent that causes bacteraemia in healthy wild rabbits in Alsace, France [4]. To the best of our knowledge there is only one case of infection with this species in humans, i.e, an endocarditis in a 74-year-old man who was in close contact with rabbits [2]. In the present study, we report the second human case of B. alsatica endocarditis in a French patient in close contact with rabbits, diagnosed using specific serogical methods, including microimmunofluorescence and Western blot with cross-adsorption studies.

Detection and identification of Bartonella sp. in fleas from carnivorous mammals in Andalusia, Spain

A total of 559 fleas representing four species (Pulex irritans, Ctenocephalides felis, Ctenocephalides canis and Spilopsyllus cuniculi) collected on carnivores (five Iberian lynx Lynx pardinus, six European wildcat Felis silvestris, 10 common genet Genetta genetta, three Eurasian badger Meles meles, 22 red fox Vulpes vulpes, 87 dogs and 23 cats) in Andalusia, southern Spain, were distributed in 156 pools of monospecific flea from each carnivore, and tested for Bartonella infection in an assay based on polymerase chain reaction (PCR) amplification of the 16 S-23 S rRNA intergenic spacer region. Twenty-one samples (13.5%) were positive and the sequence data showed the presence of four different Bartonella species. Bartonella henselae was detected in nine pools of Ctenocephalides felis from cats and dogs and in three pools of Ctenocephalides canis from cats; Bartonella clarridgeiae in Ctenocephalides felis from a cat, and Bartonella alsatica in Spilopsyllus cuniculi from a wildcat. DNA of Bartonella sp., closely related to Bartonella rochalimae, was found in seven pools of Pulex irritans from foxes. This is the first detection of B. alsatica and Bartonella sp. in the Iberian Peninsula. All of these Bartonella species have been implicated as agents of human diseases. The present survey confirms that carnivores are major reservoirs for Bartonella spp.

Bartonella alsatica sp. nov., a new Bartonella species isolated from the blood of wild rabbits.

Bartonella species are considered as emerging human pathogens, with at least six different species pathogenic or possibly pathogenic for humans. However, little is known about Bartonella distribution, species polymorphism and pathogenicity in mammalian species. The objective of this work was to determine the presence, the frequency and the distribution of Bartonella species in wild rabbits (Oryctolagus cuniculus) caught in warrens in Alsace, France. Humans may come into contact with wild rabbits when hunting, especially when they are picked up with bare hands and at time of evisceration. Of 30 blood samples collected and cultured from wild rabbits, nine (30%) were positive for organisms morphologically similar to Bartonella spp. The bacteria appeared as small, fastidious, aerobic, oxidase-negative, Gram-negative rods which could be localized within erythrocytes. Their biochemical properties were similar to those of the genus Bartonella. The sequence of the 16S rRNA gene obtained from the rabbit isolates was highly related to the sequences of the different Bartonella species (97.8-99.3% similarity). The high DNA hybridization rate (81-90% similarity) between the three strains isolated from rabbit blood confirmed that they belong to the same bacterial species. Hybridization values, obtained with the nuclease-TCA method, when testing type strains of recognized Bartonella species (9-14% similarity), support the creation of a new species for the rabbit isolates. The name Bartonella alsatica is proposed for these strains isolated from the blood of wild rabbits. The type strain is IBS 382T (= CIP 105477T).